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03-101
Sigma-AldrichRIPAb+ PABPC1 - RIP Validated Antibody and Primer Set
This RIPAb+ PABPC1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This RIPAb+ PABPC1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
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RIPAb+ PABPC1 - RIP Validated Antibody and Primer Set
Overview
RIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully coprecipitating a specific RNA target (where such a specific target is known). The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. Each set also includes a negative control antibody to ensure specificity of the RIP reaction. The RIPAb+ PABPC1 set includes the PABPC1 antibody, a negative control antibody (purified mouse IgG), and positive qPCR primers which amplify a 100 bp fragment of the human ACTB cDNA. The PABPC1 and negative control antibodies are supplied in a scalable "per RIP" reaction size and can be used to functionally validate the precipitation of PABPC1-associated RNAs.
References
Product Information
Format
Purified
Control
Included negative control mouse IgG antibody and control primers specific for actin B.
Presentation
Anti-PABPC1, clone 10E10 (mouse monoclonal IgG) One vial 50 μg of protein A purified mouse IgG2b in 50 μL of PBS containing 0.1% sodium azide. Store at -20°C
Normal Mouse IgG One vial containing 125 μg purified mouse IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C.
RIP Primers, ACTB One vial containing 75 μL of 5 μM of each primer specific for actin B (ACTB). Store at -20°C. FOR: TTG TTA CAG GAA GTC CCT TGC C REV: ATG CTA TCA CCT CCC CTG TGT G
This RIPAb+ PABPC1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
RNA Binding Protein Immunoprecipitation (RIP)
Immunoprecipitation
Western Blotting
Application Notes
Immunoprecipitation from RIP lysate: RIP lysate from HeLa cells (2 X 106 cell equivalents per IP) was subjected to immunoprecipitation using 0.5 μg of either a normal mouse IgG or Anti-PABPC1 antibody. Precipitated proteins were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-PABPC1 (1.0 μg/mL). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232) (Please see figures).
Western Blot Analysis: 0.5-2 μg/mL of a previous lot detected PABPC1 in RIPA lysates from HeLa and Jurkat cells (Please see figures). Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-PABPC1 (0.5 μg/mL). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.
Biological Information
Immunogen
The PABPC1 monoclonal antibody is made against a fusion protein corresponding to full-length of human cytoplasmic poly (A)-binding protein (PABPC1).
The poly(A)-binding protein (PABP), which is found complexed to the 3-prime poly(A) tail of eukaryotic mRNA, is required for poly(A) shortening and translation initiation. In humans, the PABPs comprise a small nuclear isoform and a conserved gene family that displays at least 3 functional proteins: PABP1 (PABPC1), inducible PABP (iPABP, or PABPC4; MIM 603407), and PABP3 (PABPC3; MIM 604680). In addition, there are at least 4 pseudogenes, PABPCP1 to PABPCP4.[supplied by OMIM]
FUNCTION: SwissProt: P11940 # Binds the poly(A) tail of mRNA. May be involved in cytoplasmic regulatory processes of mRNA metabolism such as pre- mRNA splicing. Its function in translational initiation regulation can either be enhanced by PAIP1 or repressed by PAIP2. Can probably bind to cytoplasmic RNA sequences other than poly(A) in vivo. May be involved in translationally coupled mRNA turnover. Implicated with other RNA-binding proteins in the cytoplasmic deadenylation/translational and decay interplay of the FOS mRNA mediated by the major coding-region determinant of instability (mCRD) domain. SIZE: 636 amino acids; 70671 Da SUBUNIT: Component of a multi subunit autoregulatory ribonucleoprotein complex (ARC), at least composed of IGF2BP1, PABPC1 and CSDE1. Interacts with IGF2BP1. Part of a complex associated with the FOS mCRD domain and consisting of HNRPD, SYNCRIP, PAIP1 and CSDE1/UNR. Interacts with the PABPC1- interacting motif-1 (PAM1) and -2 (PAM2) of PAIP1 and PAIP2. Interacts with PAIP1 with a 1:1 stoichiometry and with PAIP2 with a 1:2 stoichiometry. The interaction with CSDE1 is direct and RNA- independent. Found in a mRNP complex with YBX2. Identified in the spliceosome C complex, at least composed of AQR, ASCC3L1, C19orf29, CDC40, CDC5L, CRNKL1, DDX23, DDX41, DDX48, DDX5, DGCR14, DHX35, DHX38, DHX8, EFTUD2, FRG1, GPATC1, HNRPA1, HNRPA2B1, HNRPA3, HNRPC, HNRPF, HNRPH1, HNRPK, HNRPM, HNRPR, HNRPU, KIAA1160, KIAA1604, LSM2, LSM3, MAGOH, MORG1, PABPC1, PLRG1, PNN, PPIE, PPIL1, PPIL3, PPWD1, PRPF19, PRPF4B, PRPF6, PRPF8, RALY, RBM22, RBM8A, RBMX, SART1, SF3A1, SF3A2, SF3A3, SF3B1, SF3B2, SF3B3, SFRS1, SKIV2L2, SNRPA1, SNRPB, SNRPB2, SNRPD1, SNRPD2, SNRPD3, SNRPE, SNRPF, SNRPG, SNW1, SRRM1, SRRM2, SYF2, SYNCRIP, TFIP11, THOC4, U2AF1, WDR57, XAB2 and ZCCHC8 (By similarity). SUBCELLULAR LOCATION: Cytoplasm. Nucleus. Note=Shuttles between the cytoplasm and the nucleus. TISSUE SPECIFICITY: Ubiquitous. DOMAIN: "SwissProt: P11940 The RNA-binding domains RRM1 and RRM2 and the C-terminus (last 138 amino acids) regions interact with the PABPC1- interacting motif-1 (PAM1) and -2 (PAM2) of PAIP1, respectively. & The RNA-binding domains RRM2 and RRM3 and the C-terminus (last 138 amino acids) regions interact with the PABPC1- interacting motif-1 (PAM1) and -2 (PAM2) of PAIP2, respectively. PTM: Methylated by CARM1. & Arg-493 is dimethylated, probably to asymmetric dimethylarginine. SIMILARITY: Contains 1 PABC domain. & Contains 4 RRM (RNA recognition motif) domains.
Molecular Weight
73 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
RNA Binding Protein Immunoprecipitation: RIP Lysate prepared from HeLa cells (2 X 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG or the anti-PABPC1 antibody and the Magna RIP Kit (Cat. # 17-700). Successful immunoprecipitation of PABPC1-associated RNA was verified by qRT-PCR using RIP Primers, ACTB (Please see figures). Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Packaging Information
Material Size
10 assays
Material Package
10 assays per set. Recommended use: ~5 μg antibody per RIP (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
03-101
04053252320415
Documentation
RIPAb+ PABPC1 - RIP Validated Antibody and Primer Set Analysenzertifikate
